PROJECT SUMMARY/ABSTRACT Opioid misuse and abuse is an epidemic of global concern to public health and safety, causing over 60,000 deaths annually in the United States. Obtaining real-time data on the misuse/abuse of opioids is a major challenge as this information is limited to population surveys and drug surveillance data. There is an urgency to develop smart city tools that will measure and monitor illicit drug metabolites to achieve crucial insight into the prevalence of opioid misuse/abuse without stigmatizing communities. Monitoring opioid metabolites in wastewater is a proven strategy to identify areas needing intervention instead of relying on emergency-room statistics and overdose deaths. However, current wastewater drug analysis requires costly laboratory-based tests that involves labor intensive sample collection, transportation, and analysis, resulting in long (days) turnaround times. Giner proposes to develop a compact, label-free Graphene Field Effect Transistor (G-FET) sensor utilizing high-specificity aptamers for rapid, accurate, and cost-effective monitoring of illicit drugs and their metabolites in wastewater samples. Since it generates rapid results compared to incumbent methods which rely on batch sampling of wastewater streams, and expensive, time consuming analysis, this technology will provide actionable real time data at a fraction of the cost of incumbent methods, encouraging broad use. The goal of Phase I program is to demonstrate a proof-of-concept test for two major street heroine metabolites in wastewater at pg/mL levels: Morphine and Norfentanyl. Multiple samples from wastewater treatment plants will be analyzed with Giner?s assay and results will be cross-validated using the gold standard HPLC-MS/MS method. The Specific Aims of the Phase I effort incide: 1) Development of a wastewater G-FET assay for Morphine and Norfentanyl; 2) Validation of sensor sensitivity and specificity; and 3) Sensor performance testing in real wastewater samples. Once developed, the technology will also be suitable for detection of any target illicit drug for which an aptamer can be prepared.